chip grade h3k4me3 antibody  (Cell Signaling Technology Inc)

Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: 10-week combined exercise preconditioning prevented 1-week hindlimb immobilization-induced muscle atrophy in mice. (A–C) Body weight, gross weight gain and food intake data during 10-week exercise preconditioning. ∗ p ​< ​0.05 vs. C; # p ​< ​0.05 vs. E. Two-way ANOVA was used and data are shown as means ​± ​standard error of the mean ( SEM ) (C, n ​= ​16, E, n ​= ​8, E+5003, n ​= ​8). (D) Food intake data during 1-week immobilization. ∗ p ​< ​0.05 vs. C. Two-way ANOVA was used and data are shown as means ​± ​ SEM . (E–F) aDMA level indicated for Prmt1 enzymatic activity after 10-week TC-E−5003 administration via subcutaneous injection at a dose of 2 ​mg/kg body weight, once daily, 5 days a week. (G–J) Skeletal muscle functions tests, G, grip strength, H, suspension time of hang test, I, time of the latency to fall in rotarod test, J, maximum voluntary climbing capacity (MVCC) test. (K–P) Total hindlimb mass and specific muscle mass of various parts of hindlimb, GAS, gastrocnemius, SOL, soleus, QUA, quadriceps femoris, lateralis, TA, tibialis anterior, EDL, extensor digitorum longus. H-S, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; # p ​< ​0.05 vs. Im; ## p ​< ​0.01 vs. Im; $ p ​< ​0.05 vs. E ​+ ​Im; $$ p ​< ​0.01 vs. E ​+ ​Im. Two-way ANOVA was used and data are shown as means ​± ​ SEM (C, n ​= ​8, Im, n ​= ​8, E ​+ ​Im, n ​= ​8, E+5003+Im, n ​= ​8).

Article Snippet: ChIP grade antibodies to Prmt1 (1:100, Proteintech, USA) were used to perform immunoprecipitation.

Techniques: Activity Assay, Injection, Suspension

Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: 10-week combined exercise preconditioning ameliorated 1-week hindlimb immobilization-induced imbalance between synthesis and degradation of protein in skeletal muscle. (A) Representative images of hematoxylin and eosin staining (H&E) of GAS muscle cross-sections. Scale bar ​= ​50 ​μm. (B) The average CSA of GAS muscle was quantified. (C–D) Real-time PCR results of TIRM63 (MuRF1), FBXO32 (Atrogin-1), Prmt1, and Sesn1 in GAS muscle. (E–N) Western blot results of Prmt1, Sesn1, p-FoxO3a, FoxO3a, Atrogin-1, MuRF1, IGF-1, pAkt-S473, Akt, mTOR, Raptor in GAS muscle. B-N, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; # p ​< ​0.05 vs. Im; ## p ​< ​0.01 vs. Im; $ p ​< ​0.05 vs. E ​+ ​Im; $$ p ​< ​0.01 vs. E ​+ ​Im. Two-way ANOVA was used and data are shown as means ​± ​standard error of the mean (C, n ​= ​8, Im, n ​= ​8, E ​+ ​Im, n ​= ​8, E+5003+Im, n ​= ​8).

Article Snippet: ChIP grade antibodies to Prmt1 (1:100, Proteintech, USA) were used to perform immunoprecipitation.

Techniques: Staining, Real-time Polymerase Chain Reaction, Western Blot

Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: Exercise preconditioning increased Sestrin1 (Sesn1) expression by activating protein arginine methyltransferase 1 (Prmt1)-p38/activating transcription factor 2 (ATF2) signaling in mice. (A) ChIP analysis of Prmt1 binding to the ATF2 sequence of mouse Sesn1 promoter in GAS muscle. (B–D) Western blot results of p-p38-T180/182, p38, and ATF2 in GAS muscle. (E–F) Co-IP results, IP: Prmt1, GAS muscle was used. (G–H) Co-IP results, IP: ATF2, GAS muscle was used. (I–J) Co-IP results, IP: p38, GAS muscle was used. A-J, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; # p ​< ​0.05 vs. Im; ## p ​< ​0.01 vs. Im; $ p ​< ​0.05 vs. E ​+ ​Im; $$ p ​< ​0.01 vs. E ​+ ​Im. Two-way ANOVA was used and data are shown as means ​± ​standard error of the mean (C, n ​= ​8, Im, n ​= ​8, E ​+ ​Im, n ​= ​8, E+5003+Im, n ​= ​8).

Article Snippet: ChIP grade antibodies to Prmt1 (1:100, Proteintech, USA) were used to perform immunoprecipitation.

Techniques: Expressing, Binding Assay, Sequencing, Western Blot, Co-Immunoprecipitation Assay

Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: Protein arginine methyltransferase 1 (Prmt1)-Sestrin1 (Sesn1) coordinated the differentiation of C2C12 myoblasts into mature myotubes. (A–B) H&E staining of C2C12 myotubes at the 7 th day of differentiation with si-Sesn, and Ad-Sesn1 treatment. Scale bar ​= ​100 ​μm. (C–D) Western blot results of Sesn1, FoxO3a, Atrogin-1 and MuRF1 in C2C12. A-D, C, control, si-Sesn1, si-RNA of Sesn1, Ad-Sesn1, Adenoviruses overexpressing Sesn1, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; # p ​< ​0.05 vs. si-Sesn1; ## p ​< ​0.01 vs. Si-Sesn1. Two-way ANOVA was used and data are shown as means ​± ​standard error of the mean ( SEM ) ( n ​= ​6 in each group). (E–F) H&E staining of C2C12 myotubes at the 7 th day of differentiation with si-Prmt1, and Ad-Prmt1. Scale bar ​= ​100 ​μm. (G–N) Western blot results of Prmt1, p-p38-T180/182, p38, ATF2, Sesn1, FoxO3a, Atrogin-1 and MuRF1 in C2C12. E-N, C, control, si-Prmt1, si-RNA of Prmt1, Ad-Prmt1, Adenoviruses overexpressing Prmt1, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; # p ​< ​0.05 vs. si-Prmt1; ## p ​< ​0.01 vs. si-Prmt1. Two-way ANOVA was used and data are shown as means ​± ​ SEM ( n ​= ​3 in each group). (O–R) Western blot results of Prmt1, ATF2 and Sesn1 in C2C12 myoblasts, ∗ p ​< ​0.05; ∗∗ p ​< ​0.01, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group). (S–W) Western blot results of Prmt1, FoxO3a, Atrogin-1 and MuRF1 in C2C12 myoblasts, ∗ p ​< ​0.05; ∗∗ p ​< ​0.01, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group).

Article Snippet: ChIP grade antibodies to Prmt1 (1:100, Proteintech, USA) were used to perform immunoprecipitation.

Techniques: Staining, Western Blot, Control

Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: Exercise preconditioning activated AMP-Activated protein kinase α2 (AMPKα2)-transcriptional co-activator PPAR-γ co-activator-1 α (PGC-1α) through protein arginine methyltransferase 1 (Prmt1)-Sestrin1 (Sesn1) in immobilized mice. (A–C) Western blot results of p-AMPKα2, AMPKα2, and PGC-1α in GAS muscle. (D–E) Co-IP results, IP: Prmt1, GAS muscle was used. (F–G) Co-IP results, IP: PGC-1α, GAS muscle was used. (H–I) Co-IP results, IP: Sesn1, GAS muscle was used. (J–K) Co-IP results, IP: PGC-1α, GAS muscle was used. (L–M) Co-IP results, IP: AMPKα2, GAS muscle was used. A-M, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; # p ​< ​0.05 vs. Im; ## p ​< ​0.01 vs. Im; $ p ​< ​0.05 vs. E ​+ ​Im; $$ p ​< ​0.01 vs. E ​+ ​Im. Two-way ANOVA was used and data are shown as means ​± ​standard error of the mean (C, n ​= ​8, Im, n ​= ​8, E ​+ ​Im, n ​= ​8, E+5003+Im, n ​= ​8).

Article Snippet: ChIP grade antibodies to Prmt1 (1:100, Proteintech, USA) were used to perform immunoprecipitation.

Techniques: Western Blot, Co-Immunoprecipitation Assay

Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: Exercise preconditioning prevented muscle atrophy through protein arginine methyltransferase 1 (Prmt1)-Sestrin1 (Sesn1)-transcriptional co-activator PPAR-γ co-activator-1 α (PGC-1α)-mediated skeletal muscle regeneration. (A–B) Western blot results of MyoD, Myf5, MEF2 and MyoG in GAS muscle. (C–D) Co-IP results, IP: MyoD, GAS muscle was used. A-B, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; ## p ​< ​0.01 vs. Im; $ p ​< ​0.05 vs. E ​+ ​Im; $$ p ​< ​0.01 vs. E ​+ ​Im. Two-way ANOVA was used, and data are shown as means ​± ​standard error of the mean ( SEM ) (C, n ​= ​8, Im, n ​= ​8, E ​+ ​Im, n ​= ​8, E+5003+Im, n ​= ​8). (E–F) H&E staining of myotubes at each differentiation time points of C2C12 myoblasts overexpressing Sesn1. Scale bar ​= ​100 ​μm ∗ p ​< ​0.05 vs. Ad-Sesn1-, unpaired Student's t-test was used and data are shown as means ​± ​ SEM ( n ​= ​3 in each group). (G–H) Western blot results of Sesn1, Prmt1, PGC-1α, MyoD, MEF2, Myf5, and MyoG in C2C12 myoblasts at each time points of differentiation. ∗ p ​< ​0.05 vs. D0, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group). (I–J) Western blot results of PGC-1α in nucleus of C2C12 myoblasts overexpressing Sesn1 at each time points of differentiation. ∗∗ p ​< ​0.01 vs. Ad-Sesn1, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group). (K–L) Western blot results of PGC-1α in cytoplasm of C2C12 myoblasts overexpressing Sesn1 at each time points of differentiation. ∗∗ p ​< ​0.01 vs. Ad-Sesn1-, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group).

Article Snippet: ChIP grade antibodies to Prmt1 (1:100, Proteintech, USA) were used to perform immunoprecipitation.

Techniques: Western Blot, Co-Immunoprecipitation Assay, Staining

Journal: Redox Biology

Article Title: Islet regeneration protein Reg3g promotes macrophage clearance of β cell-derived dysfunctional mitochondria-rich vesicles to mitigate T2DM

doi: 10.1016/j.redox.2025.103996

Figure Lengend Snippet: Reg3g downregulates P2RX7 by promoting HSPG-NF-κB complex formation to maintain NF-κB in the cytoplasm. (A–C) The RAW264.7 cells were treated with Reg3g or PBS for 24 h in the presence of mEVs in these indicated experiments. (A) Representative NF-κB immunofluorescence (n = 3). (B) Western blot analysis of NF-κB in whole cell lysate (n = 3). (C) Western blot analysis of NF-κB in cell nucleus lysate (n = 3). (D) Predicted NF-κB binding site motif. (E) The putative NF-κB binding sites in P2rx7 promoter. The start site of transcription (TSS) is denoted +1 and the location of potential NF-κB binding site has been shown relative to TSS. (F–H) The RAW264.7 cells were treated with Reg3g in the presence of mEVs in these indicated experiments. (F) ChIP-qPCR analyzed the binding of NF-κB to P2rx7 promoter (n = 3 to 5). (G) Co-localization of HSPG and NF-κB. The line charts represent fluorescence intensity (MFI), which is presented the distance from α to ω in the images (n = 3). (H) The interaction of HSPG and NF-κB was determined by co-immunoprecipitation (n = 3). Data are presented as the means ± SEMs. For B and C, statistical significance was calculated using Student's unpaired two-tailed t -test. For F, statistical significance was calculated using ANOVA with Tukey's post hoc comparison. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: DNA was sheared enzymatically and immunoprecipitated with the specific ChIP-grade antibodies against NF-κB (#8242, CST).

Techniques: Immunofluorescence, Western Blot, Binding Assay, ChIP-qPCR, Fluorescence, Immunoprecipitation, Two Tailed Test, Comparison